Dear Mr. or Ms
We are working in a small lab from Canarias Island (Spain). We work with
goat brain with manual procedure, our problems are several in the
1. We obtain very dry blocks, when we try to cut with the microtome,
the tissues are fragile and it is impossible to obtain the correct tissue.
2. When "we get proper block" (we can cut better the tissue), then we
try to put in the float, the tissue appears like a white colour in the
slide. After when we finish the stain process (hematoxyline-eosina and
kluver barrera both of them) the tissues are not attached on the glass
I explain you in fast way our protocol:
1. Fixation: the whole brain is embedded in Formol 10% (1 week
2. Dehydration: 80º etanol 30 minutes; 96º 30 min x 2 times; 100 º 30
min x 5 times; xylene 1 hour x 2 times
3. Tissue embedding in paraffin. In cassetes with pure paraffin all
night- Only 1 step other times 2 changes in paraffin 1 hour
4. Cut with microtome at 8 um, slide with polylisine.
5. Stain process (Hematoxyline-Eosine and kluver barrera)
Thank a lot!!
Instituto de Investigación y Ciencias de Puerto del Rosario
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